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At least 865 records · Page 48Linked to original sources
PubMed1995

DNA fingerprinting in silkworm Bombyx mori using banded krait minor satellite DNA-derived probe.

The genomic DNA from thirteen different ecotypes and inbred lines of silkworm, Bombyx mori, were analyzed by digesting with BstNI and HinfI restriction enzymes followed by hybridization with banded krait minor satellite DNA (Bkm)-2(8) minisatellite probe. The DNA fingerprinting revealed 9-31 discrete intense bands, some of which were ecotype/inbred line-specific. Individual specific DNA fingerprints in two representative genotypes and their F1 hybrid offspring were also obtained. Individuals of a given parental line showed very similar profiles and the hybrid offspring showed the combined profile of both parents. The presence of bands specific to diapausing and nondiapausing strains and to particular genotypes indicate their potential use for marker-assisted breeding and varietal identification.

Animals↗
PubMed1995

Genetic self-management in a captive colony of mandrills (Mandrillus sphinx) as revealed by DNA minisatellite fingerprints.

DNA fingerprints using three oligonucleotide probes, a (GTG)5 short tandem repeat sequence, and 2 corresponding to the core sequences of minisatellites 33.15 and 33.6, have been used to generate DNA fingerprints of a captive colony of mandrills (Mandrillus sphinx) housed at the Centre International de Recherches Médicales de Franceville (CIRMF), in order to monitor paternity. A colony of 6 males and 7 females was established in a 6 ha. rain-forested enclosure in 1983. Band sharing scores indicated that none of the founders were related. By April 1994 the colony had increased to 84 individuals. Forty-nine of the 77 births were first-generation infants, a further 27 were second-generation offspring, and one was a third-generation mandrill. Five animals died during the study, and one was removed for husbandry reasons. Overall band sharing between individuals increased markedly and the power to discriminate between first- and second-generation infants was reduced; the probability of two individuals sharing the same pattern increased 1000-fold from 10(-6) to 10(-3). The dominant male in the group effectively guards access to all receptive females during the annual breeding season. Paternity analysis confirmed that the dominant male is the father of the great majority of offspring born during this term of dominance. Daughters born before 1991 to either of the two dominant males reached sexual maturity during this tenure. Incestual inbreeding occurred with both males, but only 25% of all possible cases resulted in the birth of a viable infant; one male fathered infants on 3 of 13 possible occasions and a second male on 2 of 7 occasions. Female avoidance strategies and shifts in male dominance in this closed mandrill colony appeared to be timed in a way that reduces incest.

Animals↗
PubMed1995

DNA fingerprinting in captive population of the endangered Siberian crane (Grus leucogeranus).

DNA fingerprinting was used to estimate genetic diversity within the endangered Siberian crane (Grus leucogeranus) captive population consisting of several dozens of founders originating from the two wild populations of eastern and western Siberia. Similarity and difference among captive individuals were demonstrated by the unweighted pair-group (UPGMA) clustering procedure. Quantitative characteristics of the eastern and western captive population groups such as average percentage differences (APD) and heterozygosity showed a high extent of genetic variability of 77.9-79.3% and heterozygosity of 0.85-0.72 within each group. Genetic heterogeneity of the captive population structure observed here provides guidelines for management of the species gene pool in captivity. These data also indicate that monitoring of genetic diversity through DNA fingerprinting can facilitate the efforts of Siberian crane management and restoration.

Animals↗
PubMed1995

Social and related implications of DNA fingerprinting and other new DNA technologies.

The discovery of the structure DNA at the beginning of this half of the century, and the recognition that it is virtually the universal genetic material, made it imperative for man, sooner or later, to apply this knowledge for unexpected ends. Genetic engineering, DNA fingerprinting, sequencing of whole genomes (be they of man, plants or microorganisms), or exploitation of the differences between the DNA of the male and the female (for example, of the X and the Y sperm) have thus all been historical imperatives. Today, many such predicted technologies are a reality or have come close to being so. Such DNA-based technologies have enormous social implications, discussed in this article. The impact of these technologies on different societies is likely to be different, in view of the different social conditions that prevail, and the different historical background and cultural traditions. In India, for example, the use of DNA fingerprinting could prevent the large-scale exploitation of women, and several kinds of litigation. On a global scale, the availability of this technique and its refinement to a stage where it would become virtually foolproof could act as a deterrent to certain kinds of crime. There will also be unprecedented benefits to agriculture but also new problems that will need to be faced and resolved. Wisdom demands that we prepare ourselves for a large-scale application of the new DNA technologies that are a historical imperative, understand the problems that are likely to arise, and plan to cope with them before they descend on us in full force.

Agriculture↗
PubMed1997

Informative enzyme/probe combinations for the multilocus DNA fingerprinting of marsupials.

DNA fingerprinting has become an invaluable tool in the study of population genetics, paternity success, and individual identification; however, the species specificity of some methods has made the wide-range screening of many different species very time-consuming. In this study we describe the development and application of reliable and informative DNA fingerprinting techniques in a range of marsupial species using three different restriction enzyme and two oligonucleotide probe combinations. Six species from four marsupial families, the koala (Phascolarctidae), tammar wallaby (Macropodidae), southern hairy-nosed wombat (Vombatidae), kowari, and dusky and brown marsupial mice (Dasyuridae) were examined. Restriction enzymes HinfI AluI and HaeIII were used in combination with the digoxygenin (DIG)-labelled oligonucleotide probes (CAC)5 and (GGAT)4. The combinations of HinfI/(GGAT)4, AluI/ (CAC)5 and AluI/(GGAT)4 were the most informative, providing highly resolved bands, low background, and the lowest band sharing between individuals. The genetic diversity evident within the different species showed a clear relationship between the level of band sharing and population size. The greatest levels of band sharing were found in the kowaris (80%), which were part of a long-term captive colony originating from a few founders, and the lowest levels of band sharing were found in the marsupial mice (30-35%) and tammar wallaby (45%), which were caught from large outbred wild populations.

Animals↗
PubMed1998

Oligonucleotide DNA fingerprinting optimized to determine parentage in three beetle species.

Oligonucleotide fingerprinting was performed on three beetle species from different families (Coleoptera: Staphylinidae: Aleochara curtula; Silphidae: Nicrophorus vespilloides; Tenebrionidae: Blaps lethifera) to obtain detailed information on parentage in mating systems. We report variations in the number of hybridizing fragments for the different species depending on the combinations of probes and restriction enzymes used. In addition to conventional multilocus fingerprint patterns, we established a single locus system for A. curtula (GTG5; HinfI) and described an oligolocus system in N. vespilloides (GATA4; HaeIII).

Animals↗
PubMed2003

Genetic differentiation and subspecies development of the giant panda as revealed by DNA fingerprinting.

Over the last 100 years giant pandas (Ailuropoda melanoleuca) have been separated into six completely isolated mountain ranges. DNA fingerprinting revealed different differentiation patterns in giant pandas, including early-stage, late-stage, recent divergence and recent genetic depression. A separation around 10 000 years ago resulted in highly significant differences in DNA fingerprints and morphological characteristics between Qinling and Sichuan populations. Supported by morphological differences, the genetic data were used to classify the Qinling population as a new subspecies, A. m. qinlingensis, while the Sichuan populations were classified into the original subspecies, A. m. melanoleuca. Thus, the Qinling population deserves management as a separate unit. In the Sichuan populations, two management units were defined, including Qionglai-Minshan and Daxiangling-Xiaoxiangling-Liangshan. Our data suggest urgent measures are needed to establish green corridors between subpopulations in each mountain range to increase gene flow and genetic variation to ensure long-term survival.

Animals↗
PubMed2006

Enterotoxigenic Bacillus spp. DNA fingerprint revealed in naturally contaminated nonfat dry milk powder using rep-PCR.

Dry milk powders and functional ingredients frequently contain high levels of viable bacterial spores, some of which may result in growth of toxigenic Bacillus spp. in reconstituted and temperature-abused foods. Samples from nonfat dry milk (NFDM), infant milk formula (IMF), coffee creamer, lecithin, and cocoa powder were subjected to a short heat treatment followed by enrichment in tryptone phosphate glucose yeast extract (TPGY) broth at 32 degrees C for 12-25 hours to obtain cell densities of 10(6) CFU ml(-1). DNA was extracted using a modification of established protocol, leading to the development of an optimized method for each food system. Purified DNA was amplified by rep-PCR using extragenic sequence-targeting primers and optimized for each food. PCR fingerprints from each food were analyzed electrophoretically for banding patterns earlier correlated to that of enterotoxigenic Bacillus spp. and Bacillus cereus positive control DNA fingerprints. Reverse passive latex agglutination (RPLA) and Bacillus Diarrhoeal Enterotoxin Enzyme Linked Immunosorbent Assay (Tecra Diagnostics) confirmed the presence of HBL and NHE enterotoxin production in NFDM, Coffee creamer, infant milk formula, and two lecithin samples but not in cocoa powder. These results demonstrate the utility of rep-PCR not only as a tool for bacterial genotyping, but a unique means of quality control and hygiene monitoring in food microbiology.

Animals↗
PubMed1990

Detection of genomic alterations in carcinogen-induced mouse liver tumors by DNA fingerprint analysis.

DNA fingerprint analysis was used to study structural abnormalities in the genome of mouse liver tumor cells. Liver tumors were induced in three strains of mice, namely C57BL/6J, C3H/He and B6C3F1, by a single injection of 20 micrograms/g body wt. diethylnitrosamine on day 15 after birth. DNA from liver tumors was digested with Hinfl restriction enzyme and hybridized on Southern blots with wild-type bacteriophage M13 DNA as probe. The resulting fingerprints of tumor DNA were compared with those of DNA from normal liver tissue. Genomic aberrations were detected in two out of 68 tumors analyzed, one stemming from a C57BL/6J and the other from a C3H/He mouse.

Animals↗
PubMed1999

Detection of DNA abnormalities by arbitrarily primed PCR fingerprinting: amplification of the MDM2 gene in a mediastinum fibrosarcoma.

Arbitrarily primed PCR (AP-PCR) fingerprinting method is easy and useful for analysis of genetic alterations in anonymous chromosomal regions. We applied this technology to analysis of DNA from human primary cancers and found amplification of a DNA fragment in a mediastinum fibrosarcoma. PCR-based analysis of radiation hybrid panels following cloning and nucleotide sequence determination of the fragment revealed that it was derived from a region of chromosome 12q13-q15. In this region, the MDM2 and IFNG genes were noted as known genes that could be involved in human carcinogenesis. Southern blot analysis of genomic DNA of the tumor revealed the amplification of the MDM2 gene together with the fragment locus, but not the IFNG gene. Our results demonstrated that detection of DNA alterations by AP-PCR fingerprinting without any previous knowledge of the genes and subsequent analysis of radiation hybrid panels could lead to easy identification of candidates for genes involved in carcinogenesis.

Base Sequence↗
PubMed1994

Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification.

Simple sequence repeats (SSR), or microsatellites, are ubiquitous in eukaryotic genomes. Here we demonstrate the utility of microsatellite-directed DNA fingerprinting by polymerase chain reaction (PCR) amplification of the interrepeat region. No sequencing is required to design the oligonucleotide primers. We tested primers anchored at 3' or 5' termini of the (CA)n repeats, extended into the flanking sequence by 2 to 4 nucleotide residues [3'-anchored primers: (CA)8RG, (CA)8RY, and (CA)7RTCY; and 5'-anchored primers: BDB(CA)7C, DBDA(CA)7, VHVG(TG)7 and HVH(TG)7T]. Radioactively labeled amplification products were analyzed by electrophoresis, revealing information on multiple genomic loci in a single gel lane. Complex, species-specific patterns were obtained from a variety of eukaryotic taxa. Intraspecies polymorphisms were also observed and shown to segregate as Mendelian markers. Inter-SSR PCR provides a novel fingerprinting approach applicable for taxonomic and phylogenetic comparisons and as a mapping tool in a wide range of organisms. This application of (CA)n repeats may be extended to different microsatellites and other common dispersed elements.

Animals↗
PubMed1994

Mycoplasma gallisepticum strain differentiation by arbitrary primer PCR (RAPD) fingerprinting.

We demonstrate here that the arbitrary primer polymerase-chain-reaction-based DNA fingerprinting method (also termed random amplified polymorphic DNA or RAPD) can be used to distinguish among strains of the avian pathogen Mycoplasma gallisepticum. Ten base oligonucleotide primers were used individually to prime DNA synthesis from genomic DNAs. Strain-specific arrays of DNA fragments were generated, which allowed us to identify and group isolates. Isolates of M. synoviae, M. gallinarum and M. iners yielded arrays of DNA fragments that differed markedly from those generated from the M. gallisepticum isolates using the same arbitrary primers. These results show that the RAPD fingerprinting method distinguishes genetically different strains of M. gallisepticum and indicates that it should be valuable for monitoring transmission of this pathogen.

Animals↗
PubMed2001

Improvement of RNA fingerprinting efficiency for the analysis of differential gene expression in human cardiac macro- and microvascular endothelial cells.

RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) is a powerful tool to screen differential gene expression. However, PCR-based screening techniques show a high incidence of false positive results (40-90%). In order to increase the efficiency and feasibility of RAP-PCR, the original protocol was modified and applied to analyse differential gene expression in human coronary macro- (HCEC) and microvascular (HCMEC) endothelial cells. The major modifications introduced were: (i) the use of two primers for PCR amplification, instead of reverse-transcription primer alone; (ii) the use of three cycles at low stringency followed by further amplification at high stringency; (iii) optimization of amplification cycle number, template amount, and concentration of primers, dNTP, Mg(2+); (iv) detection of fingerprints by silver staining; and (v) direct sequencing using RAP-PCR primers. Analysis of untreated and TNF alpha -stimulated (100 U ml(-1)for 1, 4, and 24 h) HCEC and HCMEC displayed 11 differentially expressed products by 18 primer combinations. Confirmation of results by RT-PCR showed that the rate of false positives attributable to our screening method was less than 20%. Among detected RAP-PCR products, the expression of Mn-superoxide dismutase, A20 zinc finger protein, and three novel genes (A/a, 4/d, 7/c) was more strongly modulated by TNF in HCEC than HCMEC. A further novel gene (B/e) was strongly expressed in HCMEC while only barely detectable in HCEC. In conclusion, modification of RAP-PCR strongly reduced the incidence of false positives, eliminated a radioactive requirement, and allowed sequencing without prior cloning, supplying an improved technology able to identify new differentially expressed genes between macro- and microvascular endothelial cells.

Cells, Cultured↗
PubMed1991

Identification of markers associated with quantitative trait loci in chickens by DNA fingerprinting.

Three approaches for identifying VNTR alleles associated with quantitative traits in chickens are described. One approach is based on the comparison of well-defined selected and non-selected control strains. The second approach is based on analyzing chickens within a breeding population ranked according to specific traits and the third approach involves segregation analysis. In this latter approach a large number of offspring of a single male segregating for a quantitative trait are produced and tested for trait association of the male DNA fingerprinting bands. In all cases pooled DNA samples of birds, rather than individual samples, are analyzed and band intensity is assumed to reflect the relative frequency of an allele. Examples from the literature and from our laboratory indicate that these methods permit the identification of DNA fingerprinting bands associated with quantitative traits. After developing locus-specific probes for these bands it should ultimately be possible to detect and map quantitative trait loci.

Animals↗
PubMed1991

Two-dimensional DNA-fingerprinting in animals.

DNA-fingerprinting has become, during the last five years, an important method of genetic analysis in medicine, veterinary medicine and other disciplines. The power of this technique, especially for genetic linkage analysis, may be enhanced in humans by using the two dimensional DNA-fingerprinting method. Here we show that this procedure can successfully be applied to different animal species, e.g. pig, dog and mouse. Optimal conditions, however, have to be determined for each species tested. With the use of marker systems as well as computer programs it will be possible to evaluate complex two-dimensional spot patterns in a short time and with high reliability.

Animals↗
PubMed1991

DNA fingerprinting: the utilization of minisatellite probes to detect a somatic mutation in the proteus syndrome.

Syndromes with localized or segmental abnormalities have been proposed to be the result of a somatic mutation leading to the presence of somatic mosaicism in the tissue. The Proteus syndrome, with its hemihypertrophy, macrodactyly and exostoses, has features which would indicate that the phenotype results from such events. The success of utilizing DNA fingerprint probes to detect somatic mutations in cancer raised the possibility that a similar approach might be successful in an investigation of two patients with the Proteus syndrome. Single band differences were detected with the probe 33.6 in a pair of monozygotic twins discordant for Proteus and in a comparison of tissue from normal and affected areas in another patient. These findings would appear to confirm the hypothesis that the Proteus syndrome results from a somatic mutation. Furthermore, the results indicate that DNA fingerprinting may offer a valuable technique for identifying probes for investigations of similar syndromes.

Abnormalities, Multiple↗
PubMed1991

Analysis of population genetic structure by DNA fingerprinting.

DNA fingerprint similarity is now being used widely to make inferences about the genetic structure of natural and domesticated populations, often with little regard to the limitations of such data. This paper provides an overview of the statistical theory of DNA fingerprint analysis with special focus on applications to natural populations for which little if anything is known about the detailed genetics of the DNA profiles. Approaches to estimating individual and population homozygosity, effective population size, population subdivision, and relatedness are reviewed, and issues concerning the biases and sampling properties of the statistics are discussed.

DNA Fingerprinting↗
PubMed1993

Paternity testing with the F10 multilocus DNA fingerprinting probe.

Empirical analysis of 200 paternity cases by multilocus DNA fingerprinting with the F10 probe showed that it was capable of distinguishing fathers from non-fathers in every case. The average exclusion probability was 0.99998. A very effective discrimination parameter was the proportion of non-maternal (test) bands which cannot be detected in the alleged father (unassignable bands) among all test bands. Values below 0.2 were seen in true fathers while in all cases of non-fathers the values were above 0.35. Minisatellite mutations occurred at a rate of 0.004 per band per child. The distribution of band-sharing among first degree relatives and unrelated individuals showed only a small overlap. Thus, band-sharing of the F10 fingerprints should provide a useful statistic for testing genetic relationships in deficiency cases.

Base Sequence↗